Carbazole derivatives for the treatment of fibrotic diseases and related symptoms, and conditions thereof

ABSTRACT

The invention provides carbazole derivatives for the treatment of fibrotic diseases (pathological collagen deposition) in tissues and organs, and related symptoms, and conditions thereof.

TECHNOLOGICAL FIELD

The present invention provides carbazole derivatives for the treatmentof fibrotic diseases (pathological collagen deposition) in essentialtissues and organs, and related symptoms, and conditions thereof.

BACKGROUND ART

-   1. Wynn T A. Cellular and molecular mechanisms of fibrosis. J.    Pathol. 2008; 214:199-210.-   2. King T E, et al, for the ASCEND Study Group. A phase 3 trial of    pirfenidone in patients with idiopathic pulmonary fibrosis. N Engl J    Med. 2014 May 29; 370(22): 2083-2092-   3. Richeldi L, et al, for the INPULSIS Trial Investigators. Efficacy    and safety of nintedanib in idiopathic pulmonary fibrosis. N Engl J    Med. 2014 May 29; 370(22): 2071-2082.-   4. McMurray J J, Packer M, Desai A S, Gong J, Lefkowitz M P, Rizkala    A R, Rouleau J L, Shi V C, Solomon S D, Swedberg K, Zile M R; for    the PARADIGM-HF Investigators and Committees. Angiotensin—neprilysin    inhibition versus enalapril in heart failure. N Engl J Med. 2014,    Sep. 11; 371(11):993-1004.-   5. Hung C, Linn G, Chow Y H, Kobayashi A, Mittelsteadt K, Altemeier    W A, Gharib S A, Schnapp L M, Duffield J S. Role of lung pericytes    and resident fibroblasts in the pathogenesis of pulmonary fibrosis.    Am J Respir Crit Care Med. 2013 Oct. 1; 188(7): 820-830.-   6. Lee R, Perry B, Heywood J, Reese C, Bonner M, Hartfield C M,    Silver R M, Visconti R P, Hoffman S, Tourkina E. Caveolin-1    regulates chemokine receptor 5-mediated contribution of bone    marrow-derived cells to dermal fibrosis. Front Pharmacol Jun. 11    2014; 5:140.-   7. Friedman S. L., Sheppard D., Duffield J. S., Violette S. Therapy    for Fibrotic Diseases: Nearing the Starting Line. Science    Translational Medicine, 2013; 5 (167): 167.

BACKGROUND

It has been estimated that 45% of all deaths in developed countries aredue to organ dysfunction resulting from fibrosis (Wynn T A et al). It isimplicated in diseases such as cardiovascular disease, pulmonaryfibrosis, liver cirrhosis, and chronic kidney failure. Virtually everyorgan in the body is vulnerable; the affected organ shrinks, hardens,and ceases to function properly.

Cellular and Molecular Mechanisms Underlying Fibrosis

Fibrosis is not a disease; it is a pathology. As such, it runsorthogonally to many different therapeutic indications and crossmultiple organs. The list of fibrosis related maladies include end-stagediseases such as: Liver cirrhosis, Kidney fibrosis, Lung-fibrosis, Heartfailure, Bone-marrow (myelofibrosis), and some also connect fibrosiswith aspects of Alzheimer's Disease.

Fibrosis can also be a part of the following conditions:Adhesion-formation following abdominal surgery, Organ fibrosis ofirradiation-damaged tissues, Over-reacting dermal scars (keloids). Itwas further shown that activated myofibroblasts play a detrimental rolein tumor progression. Tumors like breast cancer and pancreatic cancerare especially rich in those cells that in this context are known as“cancer-associated fibroblasts” (CAF).

Pathophysiology

The common denominator of all fibrosis-related diseases is thetransformation of an otherwise placid fibroblast into a “hyper-active”myofibroblast. The myofibroblast is the cell in charge of orchestratingwound repair. In response to injury, it cranks up its metabolicactivity, proliferates rapidly, secrets collagen and otherextracellular-matrix components, and develops muscle-like contractileproperties to physically close wounds. In an acute injury, like a cut ora burn, once its work done, the myofibroblast return to quiescence andreassumes its quiet life as a resident tissue fibroblast. But in injurythat is chronic and lifelong, myofibroblasts persist and eventually takeover, filling the organ with collagen, laying down scar tissue, crowdingout and eventually destroying the functional tissue in the organ. Withtime, the organ can no longer function properly; it fails.

After injury, organs in the body undergo a wound-healing response,intended to create new connective tissue. This process involves a widevariety of cells, molecules, and signaling pathways. Critical cells inthe process are the fibroblasts and the smooth muscle—likemyofibroblasts into which they differentiate which both produceextracellular matrix (ECM) and govern its resorption. Pericytes, whichtypically form a sheath around blood vessels and regulate vascularstability, are another group of fibrogenic cells, which are majorcontributors to the myofibroblast population in a mouse model ofpulmonary fibrosis (Hung C et al) Finally, epithelial-mesenchymaltransition (EMT), in which epithelial cells differentiate intomesenchymal cells, is thought to create an environment conducive tofibrosis, however, the contribution of EMT to collagen synthesis remainsdebated.

In pathological fibrosis, the balance is lost between normal productionof ECM (scar tissue) and its resorption. With progressive fibrosis, thetissue becomes less elastic and pliable and more fibrous, stringy, andtough. These changes in tissue architecture alter organ function and canultimately lead to organ failure. Pathological fibrosis in any organsystem involves more than one of these myofibroblast precursors,suggesting that combination regimens of anti-fibrotic agents, eachtargeting a different precursor, might be necessary for successfultherapy.

State of the Art Treatments of Fibrosis

Despite the overwhelming need, few agents with anti-fibrotic effectshave made it to the market, leaving millions of patients with verylimited treatment options. Clinical trials of anti-fibrotic therapiesreported beneficial results for idiopathic pulmonary fibrosis (King T E,et al; Richeldi L, et al) and heart failure (McMurray J J et al).

One of the most promising treatments focus on blocking a pathway drivenby a molecule known as TGF-B, which is as a central mediator offibrosis. One approach developed by Genzyme, Eli Lilly, and Biogen Idec,includes a variety of humanized antibodies against TGF-B. However, therehave been concerns that inhibiting this pathway could also impair thehealthy process of tumor suppression. Another strategy, developed byGilead Sciences is using antibodies to block an enzyme that promotescross-links among proteins in scar tissue. Such cross-linking is thoughtto stiffen the scar and hamper normal tissue repair and scar resolution.A third approach, explored by Genentech, Novartis, MedImmune and Sanofi,targets cytokines, or messenger chemicals, IL-4 and IL-13, whichregulate fibrosis in several animal models. Hoffman and Tourkina areinventors on an issued U.S. patent for the use of caveolin-1 scaffoldingdomain (CSD) as an anti-fibrotic treatment in mice model of Scleroderma,in which CSD substitutes for the lost caveolin-1 and prevents fibrosis(Lee R et al). New anti-fibrosis agents tested in Idiopathic PulmonaryFibrosis such as pirfenidone and nintedanib slowed a decline in fibrosishowever, they have not reversed the disease (King T E et al).

The Unmet Need for a Treatment of Fibrosis

As the mechanisms underlying fibrosis are becoming better elucidated andas more and more biomarkers of disease are being identified, theinterest in developing anti-fibrotic agents has been growing. Effortsshould be made to better target anti-fibrotic therapies by developingbiomarkers that will predict which patients might benefit from aparticular therapy.

Scleroderma, the prototype of the fibrotic disease, is characterized bydeposition of excessive collagen in the skin of the face, extremities,and trunk as well as injury to the capillaries. If an anti-fibroticagent is demonstrated to be efficacious in scleroderma, that could meanthat it holds promise for treating the fibrotic diseases of the lung,heart, liver, kidneys and other organs (Friedman S L et al). In otherwords, from a pathophysiology point of view, all the above diseasesrepresents a single entity; you cure one, you cure them all.

General Description

The inventors of the present application have found that compositionscomprising at least one active agent having the general formula (I) orany salt thereof, are suitable in uses and methods of treating ofconditions or diseases associated with excessive collagen deposition:

Wherein each of R₁-R₈ is independently selected from the groupconsisting of H, OH, SH, halogen, nitro, amino, nitrilo, nitroso,acetyl, acetamido, acylamido, alkylamino, straight or branched C₁-C₅alkyl, straight or branched C₁-C₅alkenyl, straight or branchedC₁-C₅alkynyl, amine, straight or branched C₁-C₅alkoxy, straight orbranched C₁-C₅ carboxyl, straight or branched C₁-C₅ ester, straight orbranched C₁-C₅thioxy, straight or branched C₁-C₅sulfinyl, straight orbranched C₁-C₅thionyl; R₉ is selected from straight or branched C₁-C₉alkyl, straight or branched C₁-C₉alkenyl, straight or branchedC₁-C₉alkynyl, piperazinyl, pyridinyl, piperidinyl, morpholinyl andthiomorpholinyl; R₉ is substituted with at least one quaternary amino(ammonium) group or a phophonium group.

Such compositions were also found to be of use in the treatment ofconditions or diseases associated with myofibroblasts activation.

The term “myofibroblasts activation” encompasses any physiologicalprocess wherein myofibroblast became metabolically activated. In thisprocess myofibroblast (a smooth muscle-like fibroblast) is transformedfrom dormant stage to active metabolic stage. The activation of thisprocess is typically due to a response to injury, the activated cellsproliferate, secret collagen and other extracellular-matrix components,and develop muscle-like contractile properties required to close wounds.In diseases or conditions associated with fibrosis, myofibroblastspersist, filling the organ with collagen, and eventually destroying thefunctional tissue of the organ.

When referring to “conditions or diseases associated with excessivecollagen deposition” it should be understood to include any conditionsor diseases that are directly or indirectly related to myofibroblastsactivation leading to irreversible excessive collagen deposition in anorgan or a tissue of a subject.

Without being bound by theory, the inventors have found that compositiondisclosed herein behave as a chemo-attractant of fibrocytes/fibroblasts,causing them to transfer from vital organs via the blood stream to theinjection site where the composition has been administered. Thismechanism leads to sparing the vital organs from development offibrosis. Thus, the compound of general formula (I) is an anti-fibroticagent that can be used for treatment of the fibrotic diseases of thelung, heart, liver, kidneys and other vital organs.

Without being bound by theory, it is postulated that bone-marrow derivedcirculating fibrocytes serve as precursor cells that home to the sitewhere the carbazole derivative of the invention was injected anddifferentiate there to myofibroblasts. Thus, by attracting them to the“neutral territory” of s.c. fat, they are no longer available to vitalorgans, namely, myofibroblast trap. In addition, it could also bepostulated that several factors (such as for example TGF-β3, Pentraxin2and IL-10) might be secreted during local fibrosis at the site where thecarbazole derivative of the invention was injected, and therebyinhibiting systemic fibrosis.

In some embodiments, at least one of R₁-R₈ is independently selectedfrom the group consisting of OH, SH, halogen, nitro, amino, nitrilo,nitroso, acetyl, acetamido, acylamido, alkylamino, straight or branchedC₁-C₅ alkyl, straight or branched C₁-C₅alkenyl, straight or branchedC₁-C₅alkynyl, amine, straight or branched C₁-C₅alkoxy, straight orbranched C₁-C₅ carboxyl, straight or branched C₁-C₅ ester, straight orbranched C₁-C₅thioxy, straight or branched C₁-C₅sulfinyl, straight orbranched C₁-C₅thionyl.

In other embodiments, at least one of R₁, R₂, R₃, R₅, R₆, R₇ isindependently selected from the group consisting of OH, SH, halogen,nitro, amino, nitrilo, nitroso, acetyl, acetamido, acylamido,alkylamino, straight or branched C₁-C₅ alkyl, straight or branchedC₁-C₅alkenyl, straight or branched C₁-C₅alkynyl, amine, straight orbranched C₁-C₅alkoxy, straight or branched C₁-C₅ carboxyl, straight orbranched C₁-C₅ ester, straight or branched C₁-C₅thioxy, straight orbranched C₁-C₅sulfinyl, straight or branched C₁-C₅thionyl.

In some embodiments, at least one of R₁-R₈ is different than H. In otherembodiments, at least two of R₁-R₈ is different than H. In furtherembodiments, at least three of R₁-R₈ is different than H. In furtherembodiments, at least four of R₁-R₈ is different than H.

In some embodiments at least one of R₁-R₄ is a halogen or at least oneof R₅-R₈ is a halogen. In some embodiments at least two of R₁-R₄ areindependently a halogen or at least two of R₅-R₈ are independently ahalogen. In another embodiment at least two of R₁-R₈ are independently ahalogen. In further embodiments, said halogen is Br.

In some embodiments, at least one of R₁-R₄ is OH. In furtherembodiments, at least one of R₅-R₈ is OH. In some embodiments at leasttwo of R₁-R₄ are independently OH or at least two of R₅-R₈ areindependently OH. In another embodiment at least two of R₁-R₈ areindependently OH.

In some embodiments at least one of R₁-R₄ is a nitro or at least one ofR₅-R₈ is a nitro. In some embodiments at least two of R₁-R₄ areindependently a nitro or at least two of R₅-R₈ are independently anitro. In another embodiment at least two of R₁-R₈ are independently anitro. In further embodiments, at least one of R₁-R₄ is a nitro and atleast one of R₅-R₈ is a nitro.

In some embodiments, R₉ is selected from straight or branched C₁-C₉alkyl, straight or branched C₁-C₉alkenyl, straight or branchedC₁-C₉alkynyl, piperazinyl, pyridinyl, piperidinyl, morpholinyl andthiomorpholinyl, and further R₉ is substituted (at any position of thesubstituent defined above) by at least one quaternary amino (ammonium)group.

In some other embodiments, R₉ is selected from straight or branchedC₁-C₉ alkyl, straight or branched C₁-C₉alkenyl, straight or branchedC₁-C₉alkynyl, piperazinyl, pyridinyl, piperidinyl, morpholinyl andthiomorpholinyl, and further R₉ is substituted (at any position of thesubstituent defined above) by at least one a phophonium group.

In some embodiments, R₉ is a straight or branched C₁-C₉ alkylsubstituted with at least one quaternary amino group.

In another embodiment, said at least one quaternary amino group has ageneral formula (V):

Wherein each of R′, R″ and R′″ is independently selected from a groupconsisting of straight or branched C₁-C₉ alkyl, straight or branchedC₁-C₉ alkenyl and straight or branched C₁-C₉ alkynyl.

In yet another embodiment, each of R′, R″ and R′″ is independently astraight or branched C₁-C₉ alkyl.

In some embodiments a composition according to the invention comprisesat least one active agent selected from:

-   3-(3,6-dibromo-9H-carbazol-9-yl)-N,N,N-trimethylpropan-1-aminium

-   5-(9H-carbazol-9-yl)-N,N,N-trimethylpentan-1-aminium

-   5-(2-hydroxy-9H-carbazol-9-yl)-N,N,N-trimethylpentan-1-aminium

-   5-(3,6-dibromo-9H-carbazol-9-yl)-N,N,N-trimethylpentan-1-aminium

or any salt thereof.

The term “halogen” is meant to encompass any halogen moiety selectedfrom F, Cl, Br and I.

The term “nitro” is a —NO₂ moiety.

The term “amino” refers to —NH₂, —NHR, —NRR′, wherein R, R′ and R″ areeach independently selected from straight or branched C₁-C₁₀ alkyl (alsotermed “alkylamino”), straight or branched C₂-C₁₀ alkenyl, straight orbranched C₂-C₁₀ alkynyl. The term amino also includes quarternaryammonium moiety of the form —⁺NRR′R″ wherein R, R′ and R″ are as definedherein above.

The term “nitrilo” refers to —CN,

The term “nitroso” refers to a NO moiety, including C-nitroso moieties(e.g., nitrosoalkanes —R—N═O, wherein R is selected from straight orbranched C₁-C₁₀ alkanyl, straight or branched C₂-C₁₀ alkenylene,straight or branched C₂-C₁₀ alkynylene), S-nitroso moieties(nitrosothiols; —S—N═O or —RS—N═O wherein R is selected from straight orbranched C₁-C₁₀ alkanyl, straight or branched C₂-C₁₀ alkenylene,straight or branched C₂-C₁₀ N-nitroso moieties (e.g., nitrosamines;—N═N═O, RN—N═O, —RR′N—N═O), and O-nitroso moieties (—O—N═O, —RO—N═Owherein R is selected from straight or branched C₁-C₁₀ alkanyl, straightor branched C₂-C₁₀ alkenylene, straight or branched C₂-C₁₀ alkynylene).

The term “acetyl” refers to a —C(═O)CH₃ moiety.

The terms “acetamido” and “acylamido” refers to —CH₂C(═O)NH₂ andCH₃C(═O)NH— respectively.

The term “straight or branched C₁-C₅ alkyl” and “straight or branchedC₁-C₉ alkyl” encompasses a saturated hydrocarbon chain having between 1to 5 or 1 to 9 carbon atoms.

The term “straight or branched C₁-C₅ alkenyl” and “straight or branchedC₁-C₉ alkenyl” encompasses a hydrocarbon chain having between 1 to 5 or1 to 9 carbon atoms and at least one double bond.

The term “straight or branched C₂-C₅ alkynyl” and “straight or branchedC₂-C₉ alkynyl” encompasses a hydrocarbon chain having between 1 to 5 or1 to 9 carbon atoms and at least one triple bond.

The term “straight or branched C₁-C₅ alkoxy” is meant to encompass an—OR moiety wherein R is selected from a straight or branched C₁-C₅alkyl, straight or branched C₂-C₅ alkenyl and straight or branched C₂-C₅alkynyl.

The term “straight or branched C₁-C₅ carboxyl” refers to a —R—C(═O)OHmoiety wherein R is selected from a straight or branched C₁-C₅ alkanyl,straight or branched C₂-C₅ alkenylene and straight or branched C₂-C₅alkynylene.

The term “straight or branched C₁-C₅ ester” refers to a RC(═O)O— moietywherein R is selected from a straight or branched C₁-C₅ alkyl, straightor branched C₂-C₅ alkenyl and straight or branched C₂-C₅ alkynyl.

The term “straight or branched C₁-C₅ thioxy” refers to a RS— moietywherein R is selected from a straight or branched C₁-C₅ alkyl, straightor branched C₂-C₅ alkenyl and straight or branched C₂-C₅ alkynyl.

The term “straight or branched C₁-C₅ sulfinyl” and “straight or branchedC₁-C₅ thionyl” refers to a RS(═O)— moiety wherein R is selected from astraight or branched C₁-C₅ alkyl, straight or branched C₂-C₅ alkenyl andstraight or branched C₂-C₅ alkynyl.

The term “phosphonium” refers to a —P+RR′R″ moiety wherein R, R′ and R″are each selected from a straight or branched C₁-C₁₀ alkyl, straight orbranched C₂-C₁₀ alkenyl and straight or branched C₂-C₁₀ alkynyl.

The term “piperazinyl” encompasses a moiety selected from:

The term “pyridinyl” encompasses a moiety:

The term “piperidinyl” encompasses a moiety selected from:

The term “morpholinyl” encompasses a moiety selected from:

The term “thiomorpholinyl” encompasses a moiety selected from:

The compounds of the present invention, as defined above, may have theability to crystallize in more than one form, a characteristic, which isknown as polymorphism, and it is understood that such polymorphic forms(“polymorphs”) are within the scope of formulae (I). Polymorphismgenerally can occur as a response to changes in temperature or pressureor both and can also result from variations in the crystallizationprocess. Polymorphs can be distinguished by various physicalcharacteristics known in the art such as x-ray diffraction patterns,solubility, and melting point.

As used herein, the term “substituted” refers to substitution with thenamed substituent or substituents, multiple degrees of substitutionbeing allowed unless otherwise stated.

Certain of the compounds described herein may contain one or more chiralatoms, or may otherwise be capable of existing as two enantiomers or astwo or more diastereomers. Accordingly, the compounds of this inventioninclude mixtures of enantiomers as well as purified enantiomers orenantiomerically enriched mixtures. Furthermore, the compounds of thisinvention include mixtures of diastereomers, as well as purifiedstereoisomers or diastereomerically enriched mixtures. Also includedwithin the scope of the invention are the individual isomers of thecompounds of the invention, as defined above, as well as any wholly orpartially mixtures thereof. The present invention also covers theindividual isomers of the compounds represented by the formulas above asmixtures with isomers thereof in which one or more chiral centers areinverted.

It is also noted that the compounds of the present invention may formtautomers. It is understood that all tautomers and mixtures of tautomersof the compounds of the present invention, are included within the scopeof the compounds of the present invention.

In some embodiments, said composition is a pharmaceutical composition,wherein said salt is a pharmaceutically acceptable salt.

Pharmaceutical compositions of the invention may additionally compriseany other suitable substances such as other therapeutically usefulsubstances, diagnostically useful substances, pharmaceuticallyacceptable carriers or the like.

In some embodiments a compound or composition of the invention isadministered (suitable to be administered) into an adipose tissue of asubject. In some embodiments said compound or composition of theinvention is administered directly into an adipose tissue of a subject.In other embodiments said administration is via injection. In otherembodiments, said administration is a transdermal administration. Undersuch embodiments, transdermal admonition can be achieved by anytransdermal formulation known in the art and/or via a transdermaldelivery device (for example a patch containing a compound orcomposition of the invention) at a close proximity to the adipose tissuelocation of said subject (for example the direct skin or mucosal tissuein contact with said adipose tissue).

Pharmaceutical compositions of the invention comprise a compound of thesubject invention in admixture with pharmaceutically acceptableauxiliaries, and optionally other therapeutic agents. The auxiliariesmust be “acceptable” in the sense of being compatible with the otheringredients of the composition and not deleterious to the recipientsthereof.

Pharmaceutical compositions include those suitable for oral, rectal,nasal, topical (including transdermal, buccal and sublingual), vaginalor parenteral (including subcutaneous, intramuscular, intravenous,intra-adipose tissue and intradermal) administration or administrationvia an implant. The compositions may be prepared by any method wellknown in the art of pharmacy. Such methods include the step of bringingin association compounds used in the invention or combinations thereofwith any auxiliary agent. The auxiliary agent(s), also named accessoryingredient(s), include those conventional in the art, such as carriers,fillers, binders, diluents, disintegrants, lubricants, colorants,flavouring agents, anti-oxidants, and wetting agents.

Pharmaceutical compositions suitable for oral administration may bepresented as discrete dosage units such as pills, tablets, dragees orcapsules (including softgel capsules), or as a powder or granules, or asa solution or suspension. The active ingredient may also be presented asa bolus, liquid formulation or paste. The compositions can further beprocessed into a suppository or enema for rectal administration.

The invention further includes a pharmaceutical composition, ashereinbefore described, in combination with packaging material,including instructions for the use of the composition for a use ashereinbefore described.

For parenteral administration, suitable compositions include aqueous andnon-aqueous sterile injection. The compositions may be presented inunit-dose or multi-dose containers, for example sealed vials andampoules, and may be stored in a freeze-dried (lyophilised) conditionrequiring only the addition of sterile liquid carrier, for examplewater, prior to use. For transdermal administration, e.g. gels, patchesor sprays can be contemplated. Compositions or formulations suitable forpulmonary administration e.g. by nasal inhalation include fine dusts ormists which may be generated by means of metered dose pressurizedaerosols, nebulisers or insufflators.

In some embodiments, compositions of the invention include alsocompositions where the compound of the invention is formulated in a fatemulsion formulation (i.e. formulated in conventional formulationprocesses to produce an emulation comprising at least one fat component,either from a natural or synthetic source), such as for exampleIntralipid formulation (in any concentration).

The exact dose and regimen of administration of the composition willnecessarily be dependent upon the therapeutic or nutritional effect tobe achieved and may vary with the particular formula, the route ofadministration, and the age and condition of the individual subject towhom the composition is to be administered.

The invention also includes any salt of a compound of the invention,including any pharmaceutically acceptable salt, wherein a compound ofthe invention has a net charge (either positive or negative) and atleast one counter ion (having a counter negative or positive charge) isadded thereto to form said salt. The phrase “pharmaceutically acceptablesalt(s)”, as used herein, means those salts of compounds of theinvention that are safe and effective for pharmaceutical use in mammalsand that possess the desired biological activity. Pharmaceuticallyacceptable salts include salts of acidic or basic groups present incompounds of the invention. Pharmaceutically acceptable acid additionsalts include, but are not limited to, hydrochloride, hydrobromide,hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate,isonicotinate, acetate, lactate, salicylate, citrate, tartrate,pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate,fumarate, gluconate, glucaronate, saccharate, formate, benzoate,glutamate, methanesulfonate, ethanesulfonate, benzensulfonate,p-toluenesulfonate and pamoate (i.e.,1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Salts of theinvention may also include a counter anion being a halogen anion such asfor example chloride and bromide anions. Certain compounds of theinvention can form pharmaceutically acceptable salts with various aminoacids. Suitable base salts include, but are not limited to, aluminum,calcium, lithium, magnesium, potassium, sodium, zinc, and diethanolaminesalts. For a review on pharmaceutically acceptable salts see BERGE ETAL., 66 J. PHARM. SCI. 1-19 (1977), incorporated herein by reference.

In some embodiments the composition of the invention has water contentof less than 50% of total composition weight. In some other embodimentsthe water content of a composition of the invention is less than 30% oftotal composition weight. In further embodiments the water content of acomposition of the invention is less than 10% of total compositionweight. In further embodiments, the water content of a composition ofthe invention is between about 30% to about 0%, of total compositionweight. In further embodiments, the water content of a composition ofthe invention is about 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0%,of total composition weight.

In some embodiments, a composition of the invention comprises at least0.1% by weight of said at least one active agent of total composition.In further embodiments, the composition comprises between about 0.1% toabout 10% by weight of said at least one active agent of totalcomposition. In further embodiments, the composition comprises between0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% by weight of said atleast one active agent of total composition. In further embodiments, thecomposition comprises between about 1% to about 5% by weight of said atleast one active agent of total composition. In further embodiments, thecomposition comprises about 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%by weight of said at least one active agent of total composition.

In some embodiments, a composition of the invention is formulated fortopical application or parenteral injection (such as subcutaneousinjection) to a non-essential fat tissue of said patient.

The term “non-essential fat tissue” should be understood to include anyfat tissue that is not an essential part of an organ. In someembodiments, said non-essential fat tissue is abdominal subcutaneoustissue.

In some embodiments, the conditions or diseases associated withexcessive collagen deposition or with myofibroblasts activation isfibrosis.

The term “fibrosis” as used herein should be understood to encompass anycondition or disease that are related to directly or indirectly to aprocess of pathological excess of fibrous connective tissue in an organor tissue.

Diseases or disorders associated with fibrosis are selected from:pulmonary fibrosis, cystic fibrosis, kidney fibrosis, liver cirrhosis,endomyocardial fibrosis, old myocardial fibrosis, atrial fibrosis,mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis,progressive massive fibrosis, nephrogenic systemic fibrosis, Carpaltunnel syndrome, joint fibrosis, Crohn's Disease, Keloid, Scleroderma,arthrofibrosis, Peyronie's disease, Dupuytren's contracture, adhesivecapsulitis and any symptom or condition associated therewith.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A-1C: Macroscopic view of the scars at time of 2 months sacrificeof a pig treated with RZL-012 of the invention. FIG. 1A is a view of across-section cut through the middle of the scar at the non-treated side(L), no internal scar-tissue can be observed.

FIG. 1B is an external view of the scar at the treated side (R). FIG. 1Cshows the external view of the scar at the non-treated side (L).

FIG. 2A-2C: Macroscopic view of the scars at time of 2 months sacrificeof a pig treated with Kythera. FIG. 2A is a view of a cross-section cutthrough the middle of the scar at the non-treated side (L). White scartissue can be clearly see along the cut (marked by a black rectangle).FIG. 2B is an external view of the scar at the treated side (R). FIG. 2Cshows the external view of the scar at the non-treated side (L).

FIG. 3A-3D: Scar Histology. H&E staining of cross-sections of thevarious scare tissues. ×10 objective magnification. FIG. 3A shows theright side RZL-012 treated scar histology. FIG. 3B shows the left sideRZL-012 treated scar histology. FIG. 3C shows the right side Kytheratreated scar histology. FIG. 3D shows the left side Kythera treated scarhistology. While the entire width of the scar-tissue of the RZL-012treated pig fits into the field of this magnification (FIGS. 3A-B), thescar-tissue of the Kythera treated pig is around five times wider andcannot be contained as a whole under this microscope magnification.Therefore, only small part of it can be seen in this field (FIG. 3C-D).

FIG. 4A-4D: Scar Histology. H&E staining of cross-sections of thevarious scare tissues. ×20 objective magnification. FIG. 4A shows theright side RZL-012 treated scar histology. FIG. 4B shows the left sideRZL-012 treated scar histology. FIG. 4C shows the right side Kytheratreated scar histology. FIG. 4D shows the left side Kythera treated scarhistology.

FIG. 5A-5D: Scar Histology. H&E staining of cross-sections of thevarious scare tissues. ×40 objective magnification. FIG. 5A shows theright side RZL-012 treated scar histology. FIG. 5B shows the left sideRZL-012 treated scar histology. FIG. 5C shows the right side Kytheratreated scar histology. FIG. 5D shows the left side Kythera treated scarhistology.

FIG. 6A-6C: Results of Injured arteries analysis of the unilateralureteral ligation (UUO) kidney fibrosis model of Example 2 below. FIG.6A and FIG. 6B show the Adventitia/Muscular layers ratio (in percent)for each rat in the example (FIG. 6A) and in average (FIG. 6B). FIG. 6Cshows the stained tissue sections (Masson-Trichrome) wherein themuscular layer and an outer layer, excessively rich in collagen, in thekidneys defined as “Adventitia” are visible.

DETAILED DESCRIPTION OF EMBODIMENTS Example 1 Comparative ExperimentsShowing Effect of RZL-012 of the Invention and Kybella

The objective of this study is to evaluate the remote effect on localinjection of RZL-012 of the invention(5-(3,6-dibromo-9H-carbazol-9-yl)-N,N,N-trimethylpentan-1-aminium) intothe s.c. fat, on scar formation in pigs, compared with Kybella (aninjectable formulation of Deoxycholate developed by Kythera) an agentthat bluntly destroys s.c. fat.

Materials

-   -   1. Kythera preparation (according to published formulation): an        aqueous solution of 5% Phosphatidylcholine, 4.75% Sodium        deoxycholate and 0.9% Benzyl Alcohol.    -   2. RZL-012 of the invention        (5-(3,6-dibromo-9H-carbazol-9-yl)-N,N,N-trimethylpentan-1-aminium)        of the invention preparation:        5.0% (w/v) RZL-012 in the below indicated formulation:

Raw Material % (w/w) (g/vial) RZL-012 5.6 0.28 Tween 80 10 0.5 Propyleneglycol 57 2.85 Benzyl alcohol 3 0.15 Water for Injection 24.4 1.22 Totalper vial 100 5.0 Note: RZL-012 powder contains also a counter ion andsome bound water molecules, amounting to around 10% of the weight.

Experimental Model and Procedures

Eighty kilogram pigs were injected with either Kythera or RZL-012 of theinvention preparation, into the s.c. fat (1 cm depth, 27 G needle), at10 sites, divided into two clusters, on the right flank only. Theinjection at each site was of a volume of 0.5 ml. In addition, a cutthrough the s.c. fat was made at the midline of each pig at the rightand at the left flank.

Animals were housed in the animal unit of Biotechfarm, Israel and fed ona diet weighing 2% of body weight (B.W.) per day divided to two meals,based on the last effective weekly body weighing.

After 2 months from the day of treatment, the animals were sacrificed;the scars on both sides were photographed before and after making across section in the middle of each scar. Thereafter, tissue samples ofall cross-sections were formalin-fixed and processed for H&E histologystaining.

Results

FIGS. 1 and 2 show a macroscopic view of the scars at time of 2 monthssacrifice. As can be seen, the scar at the left, non-treated side, ismuch finer in RZL-012 of the invention treated pig, compared withKythera treated pig. Also, after a cross section through the scar, awhite scar tissue is clearly evident at the Kythera treated pig(surrounded by black rectangle) while no apparent scare tissue wasobserved in the carbazole derivative of the invention treated pig. Itcan be clearly be seen that scar formed at the non-treated side of theRZL-012 treated pig is much finer than the corresponding scar of theKythera treated pig (FIG. 1C vs. FIG. 2C).

FIG. 3A-3D shows the scar histology (H&E staining of cross-sections ofthe various scare tissues; ×10 objective magnification). In agreementwith the macroscopic view, the width of fibrotic tissue on both sides ofthe RZL-012 of the invention treated pig are much thinner (FIGS. 3A-3B)than those of the Kythera treated pig (FIGS. 3C-3D). Careful measurementreveals 4-5 times difference in the scar tissue width.

FIG. 4A-4D shows the scar histology of the same preparations as in FIG.3 but at ×20 objective magnification. It can be clearly seen that thescar-tissue in RZL-012 of the invention treated side (FIGS. 4A-4B)contains less collagen fibers as compared with the scar tissue of FIGS.4C-4D.

FIG. 5A-5D shows the scar histology of the same preparations as in FIG.3 but at ×40 objective magnification. It can be clearly seen that thescar-tissue in RZL-012 of the invention treated side (FIGS. 5A-5B)contains less collagen fibers as compared with the scar tissue of FIGS.5C-5D.

Example 2: Inhibition of Kidney Fibrosis by RZL-012

Animals:

Male SD Rats, 250 g.

Induction of Kidney Fibrosis:

Overnight fasted rats were anesthetized (a mixture of Ketamine 20 mg/kgand Xylazin 2 mg/kg IP). The abdominal cavity was exposed via a midlineincision up to 0.5-1 cm before penis about 3 cm along. Then the bladderwas lifted out and epididymal fat was remoted. The left ureter wasligated at two points with 4-0 sutures. The wound was closed in layerswith the same sutures. Then animals were returned to the cages. Thisunilateral uretral ligation led to urine accumulation in the affectedkidney, resulting in massive tissue fibrosis.

Treatment:

Test (RZL-012 15 mg) and reference (Vehicle) articles were injected atDay 14 and Day 42, following UUO, subcutaneously into the dorsalbilateral inguinal fat layer and the nape-of-the-neck fat, using a 27 G½″ needle at a dose volume of 0.1 ml/site.

Tissue Processing:

8 weeks after UUO the rats were sacrificed and the kidneys were removed,fixed and processed for histological examination. The tissue sectionswere stained with Masson-Trichrome which enable a distinction betweenfibrotic tissue (staining collagen fiber in Blue) and normal viabletissue (Purple-Red) (see FIG. 6C).

Quantification of Kidney Fibrosis:

In order to quantify, in an unbiased way, the fibrotic response inRZL-treated vs. non-treated rats, the dimensions of the fibrotic layersurrounding aortic blood vessel, were measured. All aortic vessels ineach affected kidney were analyzed in the following way: The vessel wallconsists of two components, the usual Muscular layer and an outer layer,excessively rich in collagen, in these kidneys, hereby defined as“Adventitia” (FIG. 6C). The net width of the adventitia and that of theMuscular layer were calculated as follows:

${{Adventitia}\mspace{14mu} {layer}} = \frac{( {{{AM}\; 1} - {M\; 1}} ) + ( {{{AM}\; 2} - {M\; 2}} )}{2}$${{Muscular}\mspace{14mu} {layer}} = \frac{{M\; 1} + {M\; 2}}{2}$

By using the ratio between Adventitia and Muscular layers, thedifference among vessels diameter was normalized.

Results:

A total of 6 RZL-012 treated kidneys and 5 Vehicle kidneys wereanalyzed. The detailed results are depicted in FIG. 6A and thecumulative statistical analysis demonstrated a significant inhibition ofkidney fibrosis by RZL-012 (FIG. 6B)

1.-24. (canceled)
 25. A method of treating a disease or disorderassociated with fibrosis in a patient in need thereof, said methodcomprising administering to said patient a composition comprising atleast one active agent having the general formula (I) or any saltthereof:

Wherein each of R₁-R₈ is independently selected from the groupconsisting of H, OH, SH, halogen, nitro, amino, nitrilo, nitroso,acetyl, acetamido, acylamido, alkylamino, straight or branched C₁-C₅alkyl, straight or branched C₁-C₅alkenyl, straight or branchedC₁-C₅alkynyl, amine, straight or branched C₁-C₅alkoxy, straight orbranched C₁-C₅ carboxyl, straight or branched C₁-C₅ ester, straight orbranched C₁-C₅thioxy, straight or branched C₁-C₅sulfinyl, straight orbranched C₁-C₅thionyl; R₉ is selected from straight or branched C₁-C₉alkyl, straight or branched C₁-C₉alkenyl, straight or branchedC₁-C₉alkynyl, piperazinyl, pyridinyl, piperidinyl, morpholinyl andthiomorpholinyl; R₉ is substituted with at least one quaternary amino(ammonium) group or a phophonium group.
 26. A method of claim 25,wherein said composition is administered via a parenteral injection. 27.A method of claim 25, wherein said injection is administered to anon-essential fat tissue of said patient.
 28. A method of treating adisease or disorder associated with excessive collagen deposition in apatient in need thereof, said method comprising administering to saidpatient a composition comprising at least one active agent having thegeneral formula (I) or any salt thereof:

Wherein each of R₁-R₈ is independently selected from the groupconsisting of H, OH, SH, halogen, nitro, amino, nitrilo, nitroso,acetyl, acetamido, acylamido, alkylamino, straight or branched C₁-C₅alkyl, straight or branched C₁-C₅alkenyl, straight or branchedC₁-C₅alkenyl, amine, straight or branched C₁-C₅alkoxy, straight orbranched C₁-C₅ carboxyl, straight or branched C₁-C₅ ester, straight orbranched C₁-C₅thioxy, straight or branched C₁-C₅sulfinyl, straight orbranched C₁-C₅thionyl; R₉ is selected from straight or branched C₁-C₉alkyl, straight or branched C₁-C₉alkenyl, straight or branchedC₁-C₉alkynyl, piperazinyl, pyridinyl, piperidinyl, morpholinyl andthiomorpholinyl; R₉ is substituted with at least one quaternary amino(ammonium) group or a phophonium group.
 29. (canceled)
 30. A methodaccording to claim 25, wherein R₉ is a straight or branched C₁-C₉ alkylsubstituted with at least one quaternary amino group.
 31. A methodaccording to claim 25, wherein at least one of R₁-R₄ is a halogen.
 32. Amethod according to claim 25, wherein at least one of R₅-R₈ is ahalogen.
 33. A method according to claim 25, wherein at least one ofR₁-R₄ is a halogen and at least one of R₅-R₈ is a halogen.
 34. A methodaccording to claim 25, wherein at least one of R₁-R₄ is OH.
 35. A methodaccording to claim 25, wherein at least one of R₅-R₈ is OH.
 36. A methodaccording to claim 25, wherein at least one of R₁-R₄ is a nitro and atleast one of R₅-R₈ is a nitro.
 37. A method according to claim 25,wherein said at least one active agent is being selected from:3-(3,6-dibromo-9H-carbazol-9-yl)-N,N,N-trimethylpropan-1-aminium5-(9H-carbazol-9-yl)-N,N,N-trimethylpentan-1-aminium5-(2-hydroxy-9H-carbazol-9-yl)-N,N,N-trimethylpentan-1-aminium5-(3,6-dibromo-9H-carbazol-9-yl)-N,N-trimethylpentan-1-aminium or anysalt thereof.
 38. A method according to claim 25, having water contentof less than 50% of total composition weight.
 39. A method according toclaim 25, wherein the composition comprises at least 0.1% by weight ofsaid at least one active agent of total composition.
 40. A methodaccording to claim 25, formulated for topical application.
 41. A methodaccording to claim 25, formulated for parenteral injection.
 42. A methodaccording to claim 25, wherein said condition or disease is fibrosis.43. A method according to claim 25, wherein said condition is selectedfrom pulmonary fibrosis, cystic fibrosis, liver cirrhosis,endomyocardial fibrosis, old myocardial infarction, atrial fibrosis,mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis,progressive massive fibrosis, nephrogenic systemic fibrosis, Carpaltunnel syndrome, joint fibrosis, Crohn's Disease, Keloid, Scleroderma,arthrofibrosis, Peyronie's disease, Dupuytren's contracture, adhesivecapsulitis and any symptom or condition associated therewith.